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Characterizing submicron vesicles with wavelength-resolved fluorescence in flow cytometry


, : Characterizing submicron vesicles with wavelength-resolved fluorescence in flow cytometry. Cytometry 25(2): 144-155

Individual synthetic vesicles 0.1-1.0 mu-M in diameter are sized by using single-particle fluorescence emission spectra obtained in a custom sheath flow cell with an imaging spectrograph and a charge-coupled device. Data are acquired at 1 Hz, with limits of detection (3-sigma) less than 6.0 times 10-3 and 1.0 times 10-4 molecules of free sulforhodamine 101 and fluorescein, respectively, and with a spectral range for fluorescence emission collection from 350 to 800 nm (0.45 nm/pixel resolution). The system is used for small-particle population analysis by analyzing a suspension of submicron, unilamellar synthetic vesicles prepared by standard procedures with phosphatidylserine and Texas red- or fluorescein head-group-conjugated dihydropalmitoylphosphatidylethanolamine. The submicron particles are individually identified, sized, and discriminated based on single-particle fluorescence emission spectra. Excellent agreement is found between fluorescence sizing data and transmission electron microscopic measurements.

US$19.90

PMID: 8891444

DOI: 10.1002/(SICI)1097-0320(19961001)25:2<144::AID-CYTO3>3.0.CO;2-H


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